Extended Data Fig. 1: Endothelial cells instruct macrophage phenotype transition via Rspondin3.
a, Schematic of investigation of the angiocrine effects of ECs on macrophages; b, mRNA levels of Rspo3 in ECs with or without activated by LPS was measured by qPCR; n=3 samples per group (mean ± sd), two-sided unpaired Student’s t-test was determined using GraphPad Prism. ns P=0.5417. c, Alternatively activated (M2) and classically activated (M1) macrophages measured by flow cytometry from three independent experiments with n=3 samples per group (mean ± sd); Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test using GraphPad Prism with individual P values (left to right) are ***P=0.0006, ****P<0.0001, ****P<0.0001, ****P<0.0001. d, Representative flow cytometry plots from three independent repeats showing ratios of M1 (CD86hiCD206lo) and M2 (CD206hiCD86lo) populations in BMDMs treated with control medium, EC-conditioned medium (EC), or activated EC conditioned medium (Activated EC) for 24h, respectively; e, Represented overlaid flow cytometry histograms from three independent repeats showing the MFI of the anti-inflammatory markers (CD206, CD301) and pro-inflammatory markers (CD86, CD80) in BMDMs stimulated with Rspondin3, LPS alone or in combination for 24h; f, Gene expression levels of anti-inflammatory maker genes (Mrc1, Arg1, Retnla, Chil3) and pro-inflammatory marker genes (Cd86, Cxcl1, Il1b, Tnf) as measured by qPCR in BMDMs treated with Rspondin3, LPS alone or in combination for 24h from three independent experiments with n=3 samples per group (mean ± sd); Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test using GraphPad Prism with individual P values (left to right) are ****P<0.0001, ****P<0.0001, ****P<0.0001, ****P<0.0001, ****P<0.0001, ***P=0.0003, ***P=0.0001, *P=0.0126, ns: P=0.9486, ****P<0.0001, ns P=0.2351, ****P<0.0001, ns P=0.9462, ****P<0.0001, ns P=0.8783, ****P<0.0001.
